16-methyl-3alpha, 17alpha-dihydroxy steroids of the pregnane series



United States Patent o 16-METHYL-3a,1'7a-DIHYDROXY STEROIDS OF THE PREGNAN E SERIES John M. Chemerda, Metuchen, Edward W. Tristranr, Cranford, David F. Hinkley, Plainiield, and Jan Ten Broeke, Watchung, N.J., assignors to Merck & Co.,

Inc., Rahway, N.J., a corporation of New Jersey No Drawing. Filed Sept. s, 1958, Set. No. 759,127

11 Claims. c1. 260397.45)

This invention relates to novel steroid compounds and 2,98%,713 Patented Apr. 18, 1961 2 In the above formulae R is hydrogen or acyl, Q is a divalent radical of a nitrogenous ketone reagent, X is a halogen having an atomic weight between 70 and 140, R is acyl., The reactions indicated are conducted as follows:, A B-OXygenated-I6-methyl-17a-hydroxy-pregnane-1 1,20-dione Compound I) is reacted with a nitrogenous ketone reagent-in a buffered medium to produce a 3-0Xygenated-16-methyl-pregnane-ll-one having attached to the C-20-position a divalent radical of a nitrogenous ketone reagent (Compound II). This latter compound is then reacted with an alkali metal borohydride particularly to novel steroid compounds useful as intermediates in the synthesis of 16-methyl-17a-hydroxy-21e acyloxy-l,4,9( 1 1 -pregnatriene-3,20-diones, and to processes for preparing theseintermediates.

It has now been found that 16-methyl-17u-hydroxy- 21 1-acyloxy-l ,4,9( 1 1 -pregnatriene-3,20-dione compounds can be synthesized by reactions indicated as follows:

to form the corresponding 3-0Xygenated-16-methyl-11B, l7u-dihydroxy pregnane, having attached to the C-20 position a divalent radical of a nitrogenous ketone reagent (Compound III). The latter compound is then contacted with a solution of a strong acid to produce a 3-oxygenated-16-methyl 17a hydroxy-9(11)-pregnene- 20-one (Compound IV). This compound is then treated with bromine to form a 3-oxygenated 16-methyl-17ahydroxy-21-bromo-9(11)-pregnene-- 20 one (Compound V), which may be converted to the corresponding 21- iodo compound by treatment with an alkali metal iodide. The latter 21-iodo or 2l-bromo compound (Compound V) is then contacted with an alkali metal salt of a hydrocarbon carboxylic acid to produce a .3-oxygenated-16- methyl-lh-hydroxy 21 acyloxy-9(11)-pregnene-20-one (Compound VI). This compound isthen contacted with mula Compound 1v CHzO R1 CompoundVII Compound V III T an oxidizing agent to produce a '16-methyl-17a-hydroxy- 2l-acyloXy-9 1 1 -pregnene-3,20-dione Compound 7 VII) This latter compound is then dehydrogenated to produce 16-methyl-17a-hydroxy-2l-acyloxy 1,4,9(11) pregnatriene-3,'2,0-dione (Compound VIII). v

This invention is concerned with the intermediate nitrogenous ketone derivatives of 3-oxygenated-16-niethyl-17ahydroxy pregnane-lLZO-dione (Compound 2) of the forwherein R is hydrogen or acyl and Q is a divalent radical of a nitrogenous ketone reagent, nitrogenous ketone derivatives of -3-oxygenated-16 methyl-l1fi,l7m-dihydroxy pregnane-ZO-one of the formula 7 wherein R is a hydrogen or acyl and Q is a divalent radical of a nitrogenous ketone reagent, 3-oxygenated-16-methyl- 17a-hydroxy-9(l'1)-pregnene-20-one (Compound IV) 3 -oxygenated 16 methyl-21halo-17u-hydroxy-9(l1)} pregnene-ZO-one (Compound V'),- 3-oxygenated-16-meth- 41)rp e natris -imm e: p ndzy n-i o t 3-oxygenated-16-methyl 17a hydroxy pregnane-1L20- v dione (Compound I).

The ketone derivative of 3-oxygenated-l6-methyl-17uhydroxy pregnane-ll,20-dione is prepared by reactinga 3 oxygenated 16 methyl 17cc hydroxy pregnanc- 1l,20-dione with a nitrogenous ketone reagent in a buffered medium. Typical examples of such i i-oxygenated: 16 methyl 170a hydroxy pregnane 11,20 dione compounds are 16a-methyl-3,17a-dihydroxy pregnane-11,20 dione, 16B methyl 3,170 dihydroxy pregnanc- -11,20- dione and l6a-methyl-3-acetoxy-l7a-hydroxy pregnane-11,20-dione. The nitrogenous ketone reagent is of the general formula QH wherein Q is asdefined above. reagents are hydroxylamine, semicarbazide, hydrazine and substituted hydrazines. The reaction is preferably carried out under anhydrous or substantially anhydrous conditions and in a solvent for the reactants. Suitable solvents are those which are nonreactive with the ketone reagent employed, such as alcohols, amides, nitriles and ethers as, for example, methanol, ethanol, acetonitrile, dimethylacetamide and dimethylformamide or mixtures thereof. The reaction temperature is not critical and may be between about 20.to 120 C., but preferably somewhat above room temperature in order to accelerate the formation of the ketone derivatives. Typical examples of 3-oxygenated-16-methyl-l7a-hydroxy pregnane?11,20-dione derivatives which may be formed are 160: methyl 3,1704 dihydroxy 20 semicarbazido pregnane ll one, 160: methyl 3 acetoxy 17ahydroxy 20 semicarbazido pregnane 11 one, 16,6- methyl 3,17oc dihydroxy 20 semicarbazido pregnane 11 one and 1604 methyl 3,17u dihydroxy- 20-oximino pregnane-l l-one.

The 3-oxygenated-16-methyl-17a-hydroxy pregnane- 11,20-dione ketone derivative is then reacted with an alkali metal borohydride to form the ketone derivative of 3-oxygenated-16 methyl-11,6',17a-dihydroxy pregnanc- 20-one. The alkali metal borohydride may be either sodium borohydride or lithium borohydride, sodium borohydride being preferred. i'l'he reaction is preferably carried out in an inert solvent which may also contain water. Typical examples of suitable solvents are others, alcohols, amides, and the like as, for example, methanol, ethanol, ethyl ether and 'dimethylformamide. The temperature at which the reaction is carried out is not critical. It may be varied between O" 'and the reflux temperature of the solvent mixture. In a preferred embodiment of this reaction step, 16ot-methyl-3,17a-dihydroxy-20-sernicarbazido pregnane-ll-one in solution in aqueous dimethylform-amide is treated with excess sodium borohydride at about 25 C. for a period of about 3 hours to form 16amethyl-3,1 1B, l7ot-trihydroxy-2l-semicarbazido pregnanc- 20-one. The excess reducing agent is then destroyed by the addition of acetic acid and the formed 11i3-hydroxy steroid recovered by removal of a portion of the solvent and the addition of water to precipitate the product. Ordinarily, because, of the strong akalinity of the boro- Typical examples of suitable nitrogenous ketone droxy pregnane 20-one is treated with a solution of a strong acid to form a 3-oxygenated-16-methyl-17a-hydroxy-9( 1 1 -p-regnene-20-one. reaction may be any strong mineral acid, typical examples being hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid. The reaction may be etfected in a homogeneous system or, as in a preferred embodiment, in a two-phase system wherein the steroid is, for the most part, dissolved in the organic phase and the acid is present as an aqueous solution. The reaction temperature is not critical, although in order to effect more rapid dehydration it is preferred to maintain the reaction at the reflux temperature of the mixture. A preferred method for carrying out the reaction is to dissolve the steroid in an inert solvent, e.g. an aromatic hydrocarbon or a chlorinated hydrocarbon such as benzene or chloroform and treating the thus formed steroid solution with an aqueous solution of hydrochloric acid to eifect hydrolysis of the nitrogenous ketone derivative and concurrent dehydration of the 11 fi-hydroxyl group. Typical examples of products which may be formed in this reaction are 16a-methyl-3,17a-dihydroxy-9-( 1 l -pregnene- 2O one, 160: methyl 3 acetoxy 17a hydroxy- 9(11) pregnene 20 one, 165 methyl 3,17a dihydroxy 9(11) pregnene 20 one and 16B methyl- 3-acetoxyl7a-hydroxy-9 1 1)-pregnene-20-one.

The 3 oxygenated 16 methyl 17o: hydroxy- 9(ll)-pregnene-20-one was then reacted with bromine to produce 3 -oxygenatedl 6-methyl- 17 a-hydroxy-2 l-bromo- 9(ll)-pre'gnene-20-one which may then be treated with an alkali metal acylate to form the corresponding 3-oxygenated 21 acyloxy 16 methyl 17a hydroxy- 9(11)-pregnene-20one. The bromination reaction is conducted in an organic solution using a ratio of approximately one mole of steroid to one mole of bromine.

Y Solvents which may be employed in this reaction are hydride reducing agents, when a 3-acyloxy steroid is employed as the starting material in the reduction, substantial hydrolysisof the 3-a cyloxy group takes place and the corresponding 3-hydroxy compound is formed. ,This l6-methyl-3,11/3,17ot-trihydroxy pregnane-ZO-one ketone derivative may be used directly in the next reaction step or it may be converted to the corresponding 3-acylate by treatment with a suitable acylating agent, such as acetic anhydride, acetyl chloride, and the like. Typical examples of the 3-oxygenated-16-methylr11,6,17et-dihydroxy pregnane 20 -one derivatives formed in this reduction step are l6a -methyl-3,11,17a-trihydroxy-ZO-semicairbazido pregnane, 1 6B methyl 3,115,170; trihydroxy 20- semicarbazido pregnane, 16 methyl- 115,1 dihydroxy 3 acetoxy' 20 semicarbazido: pregnane, and 16a methyl 3,11[3,17a --trihydroxy'- 20 oximino pregnane.

any inert solvents or the steroid compound which was dissolved bromine. Thus a lower alkanol, a chlorinated hydrocarbon, acetic acid or mixtures of these may be used. It is preferred to catalize the bromination by the addition ofa trace amount of hydrogen bromide. Bromination may be effected at temperatures of from about 25 up to about the boiling point of the solvent, although a temperature of about 50 is preferred, the reaction being substantially complete in about 40 minutes. The crude 21-bromo corn-pound may be converted without purification to the 21-iod0 compound by treatment in solution with an alkali metal iodide, for example, potas sium iodide. Examples of the 2l-halo compounds produced by these procedures are 16ot-methyl-3,'l7u-dihydroxy 21 bromo 9(11) pregnene 20 one, 165- methyl 3,17ot dihydroxy 21 bromo 9(11) pregnene 2O one, 160: methyl 3,170: dihydroxy 21- iodo 9(11) pregnene 20 one, 160: methyl 3- acetoxy 17a hydroxy 21 bromo 9(11) pregnene- 2O one and 16m methyl 3 .-acetoxy 17a hydroxy- 2l-iodo-9(11)-pregnene-20-one.

The 2l-halo compounds described hereinabove are convertedto the corresponding 21-acylates by treatment in solution with an alkali metal acylate. Either the 21- bromo or the 2l-iodo compounds may be used directly in themetathesisreaction.

In a preferred embodiment of the acylation reaction the S-OXygenated-lG-rnethyl-17a-hydroxy-21-bromo-9 1 l -pregnene-20-one is heatin thisreaction arel6u methyl-3,17u-dihydroxy-2l-acetoxy-9( 1 1 -pregnene-20-one I and 16 {3-methyl-3 ,17a dihydroxy-21-acetoxy-9 l 1-) -pregnene-'20-one.

The acid employed in the The 16 -methyl-3, l 7a-dihydroxy 2 l-acyloxy-9 1 1 -pregnene-ZO-one is then treated with a strong oxidizing agent to produce l6-methyl-l7a-hydroxy-2l-acyloxy 9(11)- pregnene-3,20-dione. Oxidizing agents employed in the reaction are preferably hexavalent compounds of chromium either in acid or alkaline solutions. Typical examples of oxidizing agents which may be successfully used are chromium trioxide in pyridine solution, chromium trioxide in acetic acid and sodium dichromate in acetic acid. Although pyridine and acetic acid are suitable solvents, other inert organic solvents or mixtures may be employed. When the preferred oxidizing agent, potassium dichromate and acetic acid, is used the reaction is essentially'complete in about 30 minutes although longer times, up to about 3 hours are generally employed to insure complete-oxidation of the 3-hydroxy group. Following the oxidation the reaction mixture is diluted with several volumes of water, whereupon the product precipitates and may be recovered by filtration. Typical examples of the 16-methyl-17u-hydroxy-21-acyloxy-9( 1 l pregnene- 3,20-dione compounds obtained in this step are 16a-methyl-17ot-hydroxy-21-acetoxy-9 1 1)-pregnene-3,20- dione and 16B-methyl-l7u-hydroxy 21 acetoxy 9(ll)- pregnene-3,20-dione. The l6-methyl-l'Za hydroxy 21- acyloxy-9 1 1 pregnene-3,20-dione is then dehydrogenated by one of the following procedures: (1) Bromination followed by dehydrobromination. (2) Microbial dehydrogenation. v v

In the first of these procedures the l6-methyl-l7a-hydroxy-2l-'acyloxy-9(ll)-pregnene-3,20-dione is contacted with bromine to produce either the 16-methyl-17a-hydroxy 21 acyloxy 4 bromo 9(11) pregnene-3,20-

'dione or the 16-methyl-17a-hydroxyl-2l-acyloxy-2,4 di

bromo-9(11)-pregnene-3,20 dione. The brominated compounds are then contacted with a solution of a base to form 16-methyl-17a-hydroxy-21-acyloxy-4,9 1 1 -pregnadiene-3,20-dion'e or 16-methyl-17a-hydroxy-21-acyloxy- 1,4,9(11)-pregnatriene-3,20-dione. Although any base may be employed, it is preferred to use a tertiary amine such as dimethylaniline, cillidine or a mixture of one of these bases with dimethylformamide. i 7

According to the. second of these procedures, the 16- methyl-17a-hydroxy-21-acyloxy-9(l1) pre ene-3,20-dione is contacted with a growing culture of a microorganism capable of effecting dehydrogenation in the A- methanol and water gave a product melting at 227-245 C.;

In a manner similar to the method described above, the corresponding 16p-methyl-3a,l7a-dihydroxy-20-semicarbazido pregnane-ll-one is prepared.

' EXAMPLEZ I6u-m ethyl-3a,11B,17a-trihydroxy-20-semicarbazido pregnane To a solution of 10 g. of 16a-methyl-3u,17u-dihydroxy- 20-semicarbazido pregnane -1l-one' in 200 ml. of tetrahydrofuran wasadded a solution of 3.0 g. sodium borohydride in 60 ml. of water with vigorous agitation at about 25 C. Almost all of the steroid dissolved. After stirring at 25 C. for 3 hours, the excess reducing agent was destroyed by the addition of 16% aqueous acetic acid until the solution was slightly acid (approximately pH 6). The reaction. mixture was then concentrated under vacuum during which time ,400 ml. of water was introduced into the system, thereby effecting crystallization of the product. When approximately 200 .ml. of distillate had been collected, the crystalline. slurry was cooled in an ice bath for two hours to effect complete precipitation of the product. The product was then filtered, washed with water and dried under vacuum. Further crystallization of a sample of the product from methanol and water gave material melting at 244-245" C., Amax, 2280, E% 343. 312,115,l7a-trihydr0xy-20-semicarbazido pregnane is preparedin accordance with the above procedure.

A mixture of 10 g. of l6a-methyl-3oz,l1,B,l7u-trihydrOxy-ZO-semica'rbazido pregnane 60 ml. of benzcne, 40 m1. of concentrated hydrochloric acid and 80 ml. of water was prepared and stirred at the reflux temperature fora period of about 4 hours. 'The-mixture was then cooled, and diluted with 120ml. of chloroform.

; The layers of the two-phase system were then separated,

ring of the steroid e.g. Bacillus sphaericus. This can be effected by adding the steroid compound as. a solid, or as a solution in a solventisuch as dimethylformamide, under sterile conditions to a. culture of the microorganism in a' nutrient medium and agitating the resulting mixture, thereby bringing about growth of the microorganisrn and dehydrogenation of the steroid compound The following examples are included for purposesof illustration and notof limitation. I I

, i EXAMPLE 1 I 16a-methyl,-3,1 7a-dihydroxy 20-smicqrbazido I pr'egnane-ll-one 1 A mixturewas, preparedincluding l0 of 1 6a-methyl-3,17o=-dihydroxy pregnane-ll,20-dione, 10. g. of semioarbazide, 3.33 g. semicarbazide hydrochloride, mlrof dimethylformamide and 100 ml. of methanol. The reaction mixture was stirred at thereflux temperature under nitrogen for four hours and then allowed to stir at 25 C. for .an-additional 2 hoursto promote crystallization of the product. 'To 'the reaction mixture was then added 300ml. ofnwater over the course of one hour, to effect complete precipitation of the product-J The slurryof crystalline product was-stirred in an ice bath for two hours and filtered. Th'e'recovered product was washed well: onthe' filter with water until free of-chloride ion (determined 'by testing successive washings with silver nitrate-solutidn') and dried under vacuu'nr'at C. -;Fur

ther purification of the product :bycry s tallizationf'gwith the organic layer containing the product was washed with water and then dried over sodium sulfate. The washed and dried extract was then concentrated to dryness under vacuum to yield a glassy'residue comprising the product.

. The residue was dissolved in 80 ml. of hot methanol and diluted with 40 ml. of warm C.) water to induce crystallize.

. preg-nene-ZO-one is formed.

crystallization of the product. The aqueous methanol solution was then cooled and the product began to The solution was then maintained at about 5 to 10 C. for about 4 hours to completely'crystallize the product. The product was recovered by filtration and washed with 50% aqueous methanol and dried under vacuum, .M. P-. 176l78 C; A sample recrystallized from methanol-water melted at 179-480" C. 'In similar manner to the above procedure and employing 1'6/3 methyl 3oc,11/3,17oc trihydroxy-ZO-semicarbazido pregnaneas the'steroid starting material, the corresponding 16B methyl 30:,17a dihydroxy-9(11)- EXAMPLE 4 1 16d-methyl-3a,17bc-dihydr0xy-21-bromo-9(11 -pregnene-' V, 20-one' j To a solution of 10 g. or IGa-meth-ylBa,17a-dihydroxy- 9(11) -pregnene-20+one in 200ml. of chloroform, heated to 47 'C., a solution of one M"hydrogen bromide-in methanol was added with stirring. I To'theflstirred solutionv'va'sv addedy31.5 mlfof 1:29 methanol-chloroform solution of bromine (1' M) of a periodof about 40 minutes wanemmwmmg. the temperature at "C'.

After -stirring1,an additional} minutes, the mixture was auoweaiemm to roomtemperaturei dofmliof saturated A sample of loo-methylaqueous sodium bicarbonate solution was added and the mixture stirred for another minutes. The organic layer containing the 21-bromo steroid product, was separated. and the aqueous layer extracted once with 20 ml. of chloroform. The two chloroform extracts were combined and dried over sodium carbonate. The dried chloroform extract was then concentrated under vacuum to dryness, giving a gummy residue. The gummy residue was dissolved in ml. of acetone from which crystalline 16ccmethyl-3 e-17a-dihydroxy-2 1-b1'QmO'r9 1 1 pregnene-- one was attained. This product was satisfactory for use without further purification.

Employing the above described procedure and using 165 methyl-3a,17a-dihydroxy-9(ll)-pregnene-20 one as the starting material the corresponding 16B-methyl- 311,170: dihydroxy-21-bromo-9(ll)-pregnene-20-one is formed.

EXAMPLE 5 16u-methyl-3u,1 7a-dihydroxy-2l-acet0xy-9(1 1 pregnene-ZO-one A mixture of the following materials was prepared:

10 g. of l6ot-methyl-3u,17a-dihydroxy-2l-bromo-9(ll)- pregnene-ZO-one 10 g. of potassium acetate 8 g. of potassium iodide 0.1 ml. of acetic acid 200 ml. of acetone acetone filtrate containing the product was added an equal volume of water which efiected precipitation of the product. After cooling for about .16 hours to eifect complete precipitation of the crystalline product, the crystals were recovered by filtration, washed with 20 ml. aqueous acetone and dried under vacuum: -M.P. 221* 226 -C. A sample recrystallized from ethyl acetate Employing the above described procedure and utilizing 166v methyl 30;,170: d-ihydroxy 21 =bromo 9(11J)- pregnene-ZO-one as the steroid startingmlaterial, the corresponding 16,8-methyl-3a, 17a-dihydro-xy-2 l-acetoxy-9 l 1 pregnene-ZO-one is formed. 7

' EXAMPLE 6 1 wmethyl-l 7rx-hydr0xy-21 -acetoxy-9 (1 1 pregnene 3,20-diam;

A solution of 10 g. of l6a methyl-3 t,17-ot dihydroxy- 2l-acetoxy-9(11)-pregnene-20-one in 300 ml. of acetic acid and a solution of 3.07, g. of sodium dichromate di-.

hydrate in 100 ml. of acetic acid were adjusted to 25 C. and mixed together all at once. The reaction mixture was maintained at 2527 C. for about 2 hours and then.

diluted with "1600 ml. ofwater. Aftercooling in an ice bath for 30 minutes, the reaction mixture containing the precipitated product was filtered andthe product .washed on the filter with water until thewashing's were neutral. The product Wasthen dried and chromato; graphed on acid-washed alumina, the product being ohtaiilcd inthe eluate comprises a -50 mixture of-chlorosteroid starting material l6ii-methyl-3a,17a-dihydroxy-21- acetoxy-9 1 1 -pregn-ene-20-one as starting material, the

corresponding 16,8-methyl-17a-hydroxy-21-acetoxy-9(l 1 pregnene-ZO-on is formed.

EXAMPLE 7 A solution of 5.0 g. of l6ot-methyl-l7a-hydroxy-2lacetoxy-9(11)-pregnene-3,20-dicne in 10 ml. of acetic acid and m1. of chloroform was prepared and cooled to 5 C. and 1.0 ml. of a solution of 1.0 lIl-l'lYdIOgEl'l bromide and acetic acid was added. A, solution of 13.1 ml. of 1.94 M bromine in acetic acid was added dropwise during one hour, maintaining the temperature at -5 to -8 C. The reaction mixture was stirred an additional 30 minutes at -5 C. and 2.2 g. of sodium acetate in 9.0 mlrof water was added all at once to neutralizethe acid. The excess bromine was then destroyed with aqueous sodium bisulfite solution. The reaction mixture was then concentrated under vacuum to 25 ml., whereupon the product separated as an oil. The addition of 25 ml. of acetic acid effected crystallization ofthe product. About 100 ml. of water was then added slowly over a period of about 30 minutes to completely precipitate the product and the mixture cooled in an ice bath for an hour. The crystalline 2,4- dibromo 16a methyl l7a-hydroxy-21-acetoxy-9(11)- pregnene-3,20-dione was filtered, washed with water and I dried under vacuum at 35 C. MP. 196 C. with decomposition I In similar manner and using lb methyl-lh hydroxy 2l-acetoxy-9(11)-pregnene-3,20-dione as thesteroid starting material 'the corresponding 2,4-dibromo-16B- methyl 17a hydroxy-21-acetoxy-9(1l)-pregnene-3,20- dione is formed.

B. DEHYDROBROMINATION .cooled to 30 C. and added slowly 'into a well-stirred solution containing 3.0 ml. of concentrated hydrochloric acid in 97 ml. of water. Another 30 ml. of water was added to the mixture and the slurry of crystalline production was maintained at 0.5" C. for about an hour and a. half. The product was recovered by filtra tiouand washed with dimethylformamide, concentrated hydrochloric acid, water and dried overnight under vacuum. Thecrude productthus obtained was chromatographed over g; of acid-washed aluminum in a column prepared in acetone ,and then displaced with a solution of one part of chloroform. and three parts of ether. Approximately 100 ml. portions of eluate were successively collected, the desired product appearing mainly in p0rtionsl8 through 39. The product was recovered from' these combined fractions by first removing the solvent and then stirring the. residue for 3 hours at'room temperaturein 11.0.m1. of'ether and 03 ml. of water. The crystalline 16ot-methyl-l7d-hydroxy v 21-acetoxy-1,4,9(1l), pregnatriene- 3,20 dione was filtered, washed with ether and dried under vacuum, .M.P. -20 C.

Insimilar manner, and using 2,4-dibromo l6 3-methylv'17tr-hydroxy-2l-acetoxy-9( l1 )-pregnene"-v 3,20 4 dione' as Starting material, ;,the' corresponding lGB-methyl-lM-hy- 'droxy-Zl-acetoxy -f1,4,9(l1) pregnatriene-3,20..- dione is f med 7 9 EXAMPLE 8: .-3

A. BROMINATION A solution of 10 g. of 1Got-methyl-17a-hydroxy-21-acetoxy-9(11)-pregnene-3,20-dione in 14.5 ml. of acetic acid and 150 m1. of chloroformwas cooled to 55 C. and 1 ml. of 1 M hydrogen bromide in acetic acid was added. A solution of 23.2 ml. of an acetic acid solution, 1.07 M in bromine and 1.0 M in hydrogen bromide was added over a period of about 2 hours while maintaining the temperature at -55 C. to 50 C. The solution wasstirred for an additional hour at 55 C. and 4.15 g. of sodium acetate dissolved in 30 ml. of water was added with vigorous agitation.- The reaction mixture was then concentrated under vacuum, diluted with water to precipitate the product, and maintained at -5 for one hour. The ,white precipitated product, 16 methyl- 4,-bromo-17ot-hydroxy-21-aeetoxy- 9(11)-pregnene 3,20 dione, was filtered, washed with water, and dried under vacuum at 35 C. M.P. 215 with decomposition gHCl1+ 0 B. DEHYDROBROMINATION asserts l7u-hydroxy-pregnane-l1,20-dione compounds are prepared in accordance with the following procedures:

A solution of 10.22 g. of methyl iodide in 50 ml. of ether is added to 1.73 g. of magnesium in 50 ml. of ether. .To the resulting ethereal solution of methyl magnesium iodide, maintained under a nitrogen atmosphere, is added 0.045 g. of anhydrous cuprous chloride. To this mixture is added, over a period of about one hour,

during which period the reaction mixture is stirred vigorously and maintained at approximately room temperature, a solution of about 5.6 g. of 16-pregnene-3a-ol-ll, 20-di0ne 3-acetate in 175 ml. of ether. lar solid separates during this addition. The resulting mixture is heated under gentle reflux for two hours after while the reaction mixture is cooled, and 125 ml. of saturated, aqueous ammonium chloride solution is added followed by 200 ml. of ether. :The layers are separated, andthe ethereal layer is Washed with'three 50 m1. portions of'water. The washed ethereal layer is dried, and

A mixture was prepared by adding together 4.83 g. I

of the above noted 4-bromo steroid,,4.83 -g. of sodium sulfate, 1.65 of semicarbazide, "16 m1. ,of dimethylformamide and 39 ml. of chloroform. -The mixture was stirred at 15-20" C. for 2 hours and then-concentrated under vacuum while .100 ml. of water was introduced into. the flask. After cooling the mixture at 0-5 for 2 hours the product,. 1out-methyl-3-semicarbazido-17ahydroxy-21-acetoxy-4,9( 1 1 -pregnadiene-20-one, was recovered by filtration, washed with water, and dried under vacuum.

The semicarbazone product prepared according to the above procedure is converted to 16ot-methyl-17a-hydroxy- 2l-acetoxy-4,9 11)-pregnadiene-3,20-dione by the following procedure: A mixture was prepared by adding together 4g. of the above .semicarbazone, 135ml. of dimethylformamide, 32 ml. chloroform, 17.5 ml. vwater, and 4.05 ml. concentrated hydrochloric acid in 26 11 111 of water. Thereaction mixture'was heatedato reflux temperature for 1 /2 hours with vigorous stirring to form 16m-methyl-17a-hydroxy-21-acetoxy-4,9( 1 1)-preg nadiene-3,20-dione. 1;:Following the heating period the reaction mixture was cooled and the organic layer containing the product wasseparated. The aqueous'layer was further extracted with three 5 m1.-portions of chlorothe solvent evaporated in vacuo to give a brown viscous oil. The latter material is heated for 15 minutes at 60- 70 C. with a mixture of 25 ml. acetic anhydride and 25 ml. of pyridine and the acetylated product is purified by chromatography on acid-Washed alumina followed by crystallization from petroleum ether to give approximately 1.5 g. of substantially pure 16a-methyl-pregnene- 3a-ol-11,20-dione 3-acetate. v

, To asolution of 0.8 g. of 16a methyl-pregnane-3a-ol- 11,20-dione 3-acetate in ml. of methanol is added 1.5 ml. of concentrated aqueous hydrochloric acid, and the resulting solution is stirred overnight at about 25 C. The reaction solution is evaporated in vacuo at 25 C. to a small volume, and the concentrated solution is poured into 50ml. ofjice Water. The white solid which precipvacuoJI The residual'material is extracte'd with'benzene,

form. All of the organicextracts were combined and stirred vigorously with a solution of 3.62 ml. of con-- centrated hydrochloric acid in 35.5 ml. of water at the reflux temperature for about 30 minutes -to increase the yield of product. The reaction mixture was then cooled,

' with dilute aqueouspotassium hydroxide solutionuntil the benzene'layer is free of pe'rbenzo'ic acid; thebenzene combined, Washed with ,a'qu er;aus sodium bicarbonate solution and water. The combined chloroform extracts were then dried and subsequently evaporated to dryness invaeuo to yield crystalline Mot-methyl-l7a-hydroxyf2lacetoxy-4,9 1 1)-pregnadiene-3,20-dione.

' In similar manner: and using 16fl-methyl-17a-hydroxy- 21-acetoxy-9(1l) pregnene 3,20 dione .as the steroid starting material the "corresponding 1613 methyl-17othydroxy-21-acetoxy-4,9(11) pregnadiene 3,20 dione is formed. a

and the benzene extract 'is-filtered to remove insoluble material.' The benzene extracts are evaporated; to .a volume: of ml. and petroleum ether is added to the cloud point.- The resulting solution is absorbed on 660 g. of acid-washed alumina; the alumina adsorbate is' then washed with 2 .liters' of petroleum. etherwi The ad sorbate is then eluted with, 85:15 petroleum-ether mixture,.and%the .ii'rst four. liters of eluate 'is'collected, and

evaporated to dryness invacuo .to-givea mixture, of enol acetates containing 116oc-methyl-17(20)-preguene3u,20v diol-lbone 3,20-diacetate. This mixturev of enolates, weighing. approximately 14 g., is,;,dissolve.d in 50 ml. of benzeneand treated-with. an excessoiper-benzoic acid overl a- 16-hour period. The reaction mixture is shaken layeriisthen Washed with water until neutral, dried a-nd the'solventevaporated in vacuo to give a crystalline mate 1o rnethyl51'7a,2Q-epoxy-pregnane-3a,20diol l l one ;2 0;diacetate. ,The' latter material is then dissolved,

, without purification, inZOQrnl. or methanol, 120 1. of water and 10g. ofpotasaium bicarbonate, and thei'esulfl irig solution is heated at reflux under nitrogen r ra p'e iriod of 1 6 hours.- Thefmethanolyisevaporated 'fronijthe' hydrolysis'solution invacumand the residual oilis exe tfactedfrom the i gltin aqueous solution withichloro "fgrma lhe chlqrqform extract is washed with waterfto neutrality,- dried,'; and='the 'chloroform'is evaporated under reduced pressure The residual oil is jtriturated; with ether, and the crystalline material thus formed is A white granu- 1 1 crystallized from ethyl acetate-petroleum ether to give l6a-methyl-pregnane-3 a,17u-diol-1 1,20-dione.

The 16cc-methyl-pregnane-3a,17 x-diol-l1,20-dione is conveniently converted to the corresponding 3-acylate, for example, the acetate by contacting a solution of 16ccmethyl-pregnane-3a,17u-diol-11,20-dione in pyridine with acetyl chloride.

The 3-oxygenated-16/3-methyl-17a-hydroxy pregnancl1,20-dione starting materials can be prepared in accordance with the following process:

To a solution of 3ct-acetoxy-16-pregnene-11,20-dione in a mixture of tetrahydrofuran and ethyl ether is added diazomethane to produce 3rz-acetoxy-16u,Hot-methyleneazopregnane-1l,20-dione (M.P. 186-190 C.) which precipitated from solution. Heating this compound at about 180 C. in vacuo produces 3oa-acetoxy-16-methyl-l6- pregnene-11,20-dione (M.P. 165-167 C.) which upon reaction with hydrogen peroxide in the presence of sodium hydroxide in methanol solution for 18 hours at room temperature aifords l6a,17a-epoxy 3a-hydroxy-l6fi-methyl-pregnane-ll,20-dione (M.P. 178-180 C.). When this compound is treated with perchloric acid in aqueous dioxane at 30 C. for 65 hours and the resulting reaction mixture is diluted with water a mixture of 3a,17ocdihydroxy 16-methyl-15-pregnene-11,20-dione and 3a, 17u-dihydroxy-l6-rnethylene-pregnene (M.P. 158-167" C.) is precipitated and recovered by filtration. Reduc tion of this mixture with hydrogen in methanol in the presence of palladium-calcium carbonate catalyst afiords a mixture of 3a,,l7a-dihydroxy-l6a-methyl-pregnane-ll, 20-dione and 3oz,17lx-dihydroxy-16fl-methyl-pregnane-l1, 20-dione sintering at 150 C. This mixture of acetates is conveniently separated by chromatography on neutral aluminum and elution with chloroform-benzene (1 to 1) and benzene yields 16a-methyl-3a,17a-dihydroxy pregnane-ll,20 dione. This 3oi-hydroxy compound is conveniently converted to the corresponding Bet-ester by treatment with an acylating agent; e.g. acetyl chloride and pyridine to form l6fi-rnethyl-3'tr-acetoxy'17a-hydroxy pregnane-11,20-dione. a

' The 16 methyl-l7m-hydroxy 21 acyloxy -'l,4,9(11) pregnatriene-3,20-dione compounds prepared by'the process of'the present invention have utility as intermediates in the. preparation of biologically active 16-methyl steroids. More specifically, these new products may be used These compounds possess unusually high anti-inflammatory and'gluco-corticoid activity and are useful. in the treatment of rheumatoid arthritis; 1 The 16 methyl-17ot-hydroxy 21 acyloxy 1,4,9(11).-.

I 12 We claim: 1. Compounds having the general formula wherein R is a member selected from the group consisting of hydrogen and an acyl radical of a lower aliphatic carboxylic acid and Q is a divalent radical of a nitrog enous ketone reagent. i

2. Compounds having the general formula wherein R is a member selected from the group consistmg of hydrogen and an acyl radical of a lower aliphatic carboxylic acid and Q is a divalent radical of a nitrogpregnatriene-3,20-di0nes which are prepared by the'process oi -the present invention may be converted to the active anti-arthritic compounds by application of the following procedure: The lG-methyl-l7a hydroxy-21-acyloxy-1,4,9(1'1') pregnatriene,2Q-dioncs; e.g. Ida-methyl 17mhydroxy 21 acyloX y-l,4,9(11)-pregnatriene 3,20 dione or B-methyl lh-hydroxy 21 acyloxy 1,4,9(11)- pregnatriene-3,ZO-dionehare treated with hypobromous acid to produce 9a-bromo-l6-methyl-17a-hydroxy11B;

enous ketone reagent. 3. Compounds havingthe general formula wherein R is a member selected from the group consisting I rogen and an acylradical of a lower aliphatic car boxylic acid.

4. Compounds having the general formula 7 wherein R is a member, selected from he oup consisting l7o't dihydroxy 2l-acetoxy l,4v-pregnadiene 3,20-dionesv which are then reacted Withpotassium acetate in ethanol to produce l6-methyl-9( l 1') oxide-17a-hydroxy-2-1-acet-' oxy-l,4-pregrradienc-3',20-dione. These 9(11)-oxides are then reacted with hydrogen fluoride and tetrahydrofuran to produce 16-methyl-9 u-luoro-115,17a dihydroxy 21- 2 l-acetoxy-l ,4'-'pregnadiene-3,ZO-dione.

within thescope of the impended claims I v ofihydrogen and anacyl'radica'l'of a lower aliphatic car'- boxyhc acid andX is'fa halogen having an atomic weight between and'140f- 5. Compounds-havingthe general formula of hydrogen and an aeyl radical of a lower aliphatic ear- References Cited in the file of this patent UNITED STATES PATENTS Wendler et a1. June 2, 1953 Chemerda et a1. Jan. 15, 1957 Julian et a1. Apr. 23, 1957 Krsek et al. Oct. 29, 1957 Hirschmann et a1. June 3, 1958 Spero et a1. June 10, 1958 Joly et a1. Aug. 26, 1958 Herzog Sept. 30, 1958 OTHER REFERENCES Oliveto et 81.: 80, I.A.C.S., 4431 (1958). 

1. COMPOUNDS HAVING THE GENERAL FORMULA 